转化生长因子β_3对鼠颌下腺细胞分泌功能的影响  

ROLE OF TRANSFORMING GROWTH FACTOR β_3 ON AMYLASE SECRETION OF SUBMANDIBULAR GLAND CELLS IN RAT

作　　者：朱耀旻[1,2] 周青[1,2] 王玉新[1,2] 

机构地区：[1]中国医科大学附属第一临床医院整形外科,沈阳110001 [2]中国医科大学附属第一临床医院口腔颌面外科,沈阳110001

出　　处：《中国修复重建外科杂志》2004年第6期490-493,共4页Chinese Journal of Reparative and Reconstructive Surgery

基　　金：辽宁省科技攻关基金资助项目 (0 0 2 2 50 0 1 )~~

摘　　要：目的　研究转化生长因子β3(transforming growth factorβ3,TGF-β3)对鼠颌下腺细胞 (ratsubmandibular gland cells,RSGCs)分泌淀粉酶功能的影响。　方法　原代及传代培养 Wistar大鼠 RSGCs,免疫组织化学鉴定细胞来源 ,按加入不同浓度 TGF-β3(0 .5、1.0、5 .0、10 .0、2 5 .0 ng/ ml)分组 ,未加入 TGF-β3的为对照组。采用噻唑蓝 (MTT)比色法、自动生化分析仪和免疫印迹法检测 2 4、4 8、72和 96小时后 TGF-β3对细胞增殖及淀粉酶分泌功能的影响。　结果　培养的 RSGCs免疫组织化学鉴定 CK 8.13、S- 10 0蛋白染色呈阳性 ,波形丝蛋白染色呈阴性。 MTT法检测各组细胞吸光度 A值与对照组比较 ,差异无统计学意义 (P>0 .0 5 )。 72和 96小时后 ,0 .5～ 10 .0 ng/ ml TGF-β3组与对照组比较 ,细胞分泌的淀粉酶明显增加 ,差异有统计学意义 (P<0 .0 1)。免疫印迹法检测颌下腺组织及培养细胞的淀粉酶提取物形成蛋白表达一致的电泳带。　结论　 TGF-β3能促进 RSGCs的分化和淀粉酶的分泌功能 。Objective To investigate the role of transforming growth factor β_3 (TGF-β_3) on the amylase secretion of rat submandibular gland cells(RSGCs).Methods The RSGCs were cultured and identified. The expressions of CK 8.13, S-100 and Vimentin in the RSGCs were examined by immunohistochemical staining. The experimental group was divided into 5 groups according to different concentrations of TGF-β_3 (0.5, 1.0, 5.0, 10.0 and 25.0 ng/ml) and no TGF-β_3 culture was used as control group. The effects of TGF-β_3 on the cell proliferation and amylase secretion were examined at the 24th, the 48th, the 72nd and the 96th hour. MTT colorimetric method was used to estimate vital force of culture cells. Amylase protein was assayed by auto-biochemistry equipment and Western blotting.Results The RSGCs were stained positively for CK 8.13 and S-100, but negatively for Vimentin. There were no significant differences in absorbency between the experimental groups and the control group(P>0.05). Compared with the control group, TGF-β_3 at concentrations of 0.5-10.0 ng/ml significantly stimulated the amylase secretion of RSGCs after 72 and 96 hours(P<0.01). But high concentration of TGF-β_3 (25.0ng/ml) showed no stimulation. Western blotting demonstrated that the cultured RSGCs and submandibular gland had the same band of amylase electrophoresis.Conclusion TGF-β_3 can stimulate RSGCs to differentiate and to secrete amylase, but TGF-β_3 has no effect on proliferation of RSGCs.

关 键 词：转化生长因子Β3 颌下腺细胞 淀粉酶 细胞培养 

分 类 号：R363[医药卫生—病理学]